Therapeutic preparation effective against ulcers



"sing the formation of such ulcers.

Patented Dec. 12, 1944 THERAPEUTIC PREPARATION EFFECTIVE AGAINST ULCERS David J. Sandweiss, Detroit, Micln, assignor to Parke, Davis & Company, Detroit, Mich, a corporation of Michigan No Drawing. Application February 29, 1940,

7 Serial No. 321,407

1 Claim. (Cl. 167- -74) 'Ifhe invention relates to products useful in the treatment of gastroenteric ulcers or inhibit- The invention relates more particularly to products obtainable from urines.

I have found that an active substance can be separated from animal or human urines of either the male or iemale species which has the valuable property of exercising a beneficial efl'ect on gastroenteric ulcers such as peptic or duodenal ulcers. My products are characterized by the fact that they do not possess activity of the socalled anterior pituitary-like hormone, and are not toxic when administered to humans.

In obtaining the new products, one may proceed in various ways to treat the urine so as to separate out the active portion. Thus, the new active principle of the invention can be separated from the urine by methods of precipitation with organic solvents, salting out methods, or by extraction methods. The invention will be illustrated by the following examples.

Example 1 A quantity of liters of male human urine is acidified to a pH of about 4.5 by the addition of acetic acid. If the urine is not clear, it may be centrifuged at this point. 500 cc. of acetone saturated with benzoic acid are then added and the urine allowed to stand for a few hours. At the end of this time benzoic acid carrying the desired fraction with it has separated out and is filtered 08. The precipitate is taken up in liter of acetone. TA small amount of insoluble material remains undissolveddn the acetone and is the fraction effective in treating ulcer. This fraction is separated from the supernatant liquid and centrifuged if necessary. It may also be washed with acetone. I! desired, the adsorption on benzoic acid and separation 01" activity from" the adsorbate can be repeated one or more times. Upon drying, taking up the product in water, filtering from any undissolved material and evaporating to dryness, the product is obtained as a dry amorphous powder which is soluble in water, insoluble in aqueous acetone containing about 50-60% by volume of acetone, insoluble inmost organic solvents, especially undiluted neutral organic solvents. Its aqueous solutions do not re' duce Fehlings solution. Its aqueous solutions are still active after heating under conditions which destroy the anterior pituitary-like sex a low temperature and preferably in vacuo.

male mice. The product also has no estrogenic activity such as that possessed by ovarian hormones, since it does not initiate an estrual cycle in ovariectomized animals.

Example 2 200 liters of male human urine are concentrated to aboutone-tenth the original volume at A concentrate is thereby obtained which is treated with 60 liters of ethyl alcohol to throw. out a precipitate which is filteredofl' after the mixture has been allowed to stand for sometime. ,The precipitate is then taken up in a quantity of water approximately 5 times as great as the precipitate and filtered 'from any insoluble matter. The filtrate is treated with a volume of ethyl alcohol equal to the filtrate itself. The addition of the alcohol produces a precipitate which is of ethyl alcohol.

I hormone. The product of this example does not show any gonadotropic activity when assayed by ,the usual method ofinjection into immature fefiltered oilf, the substance active in treating ulcers being in the filtrate. This filtrate can be concentrated and dried to give a product substantially equivalent to that obtained by Example 1 above.

Example 3 500 liters of fresh urine from normal nonpregnant women are evaporated in =vacuo to about 45 liters, 200 liters ofacetone added and the precipitate which 'iorms is filtered oil. The crude precipitate is dissolved in 5 times its quantity of water, any insoluble matter centrifuged off and the filtrate treated with an equal volume The precipitate which forms upon adding alcohol is filtered oil to obtain a filtrate containing the factor efiective in treating or preventing ulcer. The physical, chemical and physiological properties of the product of this example are substantially those possessed by the products of Examples 1 and 2, which were obtained from male urine.

Example 4 To 1000 liters of the urine of pregnant women are added 2000 liters of alcohol. The precipitate which forms. is filtered oil. and 2000 liters of an organic water immiscible liquid, such as ethyl out. Various other methods of effecting the sep- 1 2 1 aration oi the ulcer preventing factor from anterior pituitary-like hormonecan be used. For

Example 5 100' liters or non-pregnantm'ares urine are I saturated with a salt such as sodium chloride or ammonium sulfate and the insoluble matter thereby produced is collected by centriiugation.

1 example, the precipitate obtained by the addition tained from pregnancy urine. I also prefer to use urine from subjects who are not sufierin from gastric or duodenal or like ulcers, because I have found that urines from subjects thus sufferthe examples.

I V in the urine than benzoic acid, such as stearic The precipitate is washed thoroughly with a saturated solution or the salt andthen dissolved in distilled water. The salt may be removed mm the solution .-by dialysis or'the active fraction can I be precipitated out or the aqueous solution by the use of organic solvents or by other means such'a's those describedin. the above examples.

The-products of thisinvention and 01' the above examples are characterized by their very pronounced effect or reducing gastric secretion and acidity. Heating, tor-"example, under the conditions mentioned above for destruction of any fraction having-anterior pituitary-like activity, does not destroythis' property of the new products 0t reducingthe amountioi secretion and the acidity of gastric juice. Heating under the same conditions also does not destroy the property of being eilectiveagainst ulcers.

The products of this inventionmay be administered to-numans parenterally, preferably subcutaneou'sly ,or intramusculai'ly, in accordance with the practice known to those familiar to this 'rne invention is not limited tense of the urines described in the examples-as starting materials for obtaining the new, active principles. Other urines o1 mammals, both pregnant and non-preg name-may be used. However, I preier the use or non-pregnant female urine. or male urine as startacid, cinnamic acid, naphthoic acid, adipic acid,

etc. Various inorganic precipitates, such as molybdic or tungstic acid or phosphotungstic acid may be used to remove the activity from the main body of the urine source material. The active fraction can be removed from such acid substances, either by a solvent which dissolves the acid but does not dissolve the active fraction, for example as described above with benzoic acid, or by adding a basic substance to the acid which is capable of forming a salt of the acid less soluble than the acid itself, so that the acid can bev removed from the solution while the active fraction remains.

The foregoing description of the invention shows that the new active principle can be obtained by any method which causes separation from mammalian urine of normally water-soluble constituents which are insoluble in organic solvents such as alcohol and acetone.

What I claim as my invention is:

A producteflect'ive against ulcers, which is derived from urine of mammals not suffering from a gastric or duodenal ulcer, havingthe following properties-not toxic to humans, showing no gonadotropic activity on immature female mice,

"- ing materials, since it is" not necessary to separate out anterior pituitary like hormones which may be present with theactive i'raction' when it is ob- 1 solvents, and the aqueous solutions of which are not inactivated by moderate heating.

' DAVID J. SANDWEISS. 

